A Comprehensive Screening and Identification of Genistin Metabolites in Rats Based on Multiple Metabolite Templates Combined with UHPLC-HRMS Analysis

Abstract

Genistin, an isoflavone belonging to the phytoestrogen family, has been reported to possess various therapeutic effects. In the present study, the genistin metabolites in rats were investigated by UHPLC-LTQ-Orbitrap mass spectrometer in both positive and negative ion modes. Firstly, the data sets were obtained based on data-dependent acquisition method and then 10 metabolite templates were established based on the previous reports. Then diagnostic product ions (DPIs) and neutral loss fragments (NLFs) were proposed to efficiently screen and ascertain the major-to-trace genistin metabolites. Meanwhile, the calculated Clog P values were used to identify the positional isomers with different retention times. Consequently, a total of 64 metabolites, including prototype drug, were positively or putatively characterized. Among them, 40 metabolites were found according to the templates of genistin and genistein, which was the same as the previous research method. After using other metabolite templates, 24 metabolites were added. The results demonstrated that genistin mainly underwent methylation, hydrogenation, hydroxylation, glucosylation, glucuronidation, sulfonation, acetylation, ring-cleavage and their composite reactions in vivo biotransformation. In conclusion, the research not only revealed the genistein metabolites and metabolic pathways in vivo comprehensively, but also proposed a method based on multiple metabolite templates to screen and identify metabolites of other natural compounds.

A Complete Study of Farrerol Metabolites Produced In Vivo and In Vitro

“…(3) Metabolic interpretation: accurate mass datasets, parent drug cleavage patterns, and related drug biotransformation information were used to describe metabolic processes [23]. (4) As an important parameter, cLog P values predicted by ChemDraw 14.0 were used to discriminate between metabolite isomers [25,26], as metabolites with larger cLog P values are usually eluted more slowly in reverse-phase chromatographic systems [23].…”

“…Drug metabolism is important for the study of drug safety and toxicity, and therefore plays a vital role in drug discovery and clinical use [18]. The use of rats for in vivo metabolism investigations has been proven to be feasible and efficient [24,25,26], while liver microsomes are well suited for metabolite identification, metabolite preparation, and toxicity research in vitro [19,27]. Herein, 42 farrerol in vivo metabolites and 15 farrerol in vitro metabolites were identified.…”

A Rapid High Throughput Vibration and Vortex-Assisted Matrix Solid Phase Dispersion for Simultaneous Extraction of Four Isoflavones for Quality Evaluation of Semen Sojae Praeparatum

“…Among them, genistein is an effective anticancer substance, which could prevent, delay or block the occurrence of cancer through its multi-effect mechanism ( Russo et al, 2016 ). Genistin has a variety of therapeutic effects, such as reducing the risk of osteoporosis and postmenopausal symptoms, anti-inflammatory, anticancer, heart protection, and antioxidant, etc ( Liang et al, 2018 ; Islam et al, 2020 ). Daidzin could stimulate glucose uptake and has a hypoglycemic effect ( Meezan et al, 2005 ).…”

Chemical Constituent Profiling of Paecilomyces cicadae Liquid Fermentation for Astragli Radix

“…In CID mode, compounds are often divided into two parts, such as product ions (emerge in ESI-MS/MS spectra due to their property of being easily ionized) and neutral fragments (observed in ESI-MS/MS spectra due to their mass difference and neutral characteristics) [18,19], which are complementary in structure. It is well documented that compounds with similar backbone structure exhibit comparable fragmentation patterns, resulting in certain diagnostic product ions (DPIs) and regular neutral loss fragments (NLFs).…”